摘要
根据果蝇的ＣＹＰ４Ｄ１保守区氨基酸序列，设计了１对简并引物，从致倦库蚊溴氰菊酯抗性株的基因组ＤＮＡ中，扩增出１条特异性ＤＮＡ片段，其大小与设计的细胞色素Ｐ４５０基因片段相符。将这个片段与ｐＵＣ１９质粒重组，并克隆到ＤＨ５α大肠杆菌中，经筛选后测序，得到１条长４５６ｂｐ的核酸序列。将推导的氨基酸序列进行同源性分析，表明该序列与ＣＹＰ４家族成员同源性较高，提示该序列属于ＣＹＰ４家族；该序列与ＣＹＰ４Ｊ亚家族中目前仅知的３个成员的同源性分别为５１．５６％、５５．４９％、５６．２５％，进一步提示该序列可能为ＣＹＰ４Ｊ亚家族中的一员。
A pair of degenerate primers according to the conservative amino acid sequences of CYP4D1 in Drosophila melanogaster was designed and used to amplify cytochrome P450 from genomic DNA of deltamethrinresistant strain of C.quinquefasciatus by PCR technique.A DNA fragment sized as designed was obtained.The fragment was recombinated with pUC19,and cloned into DH5α.A positive clone was selected to sequence,and a nucleotide sequence of 456bp was obtained.Homologous analysis on the deduced amino acid sequence showed that the sequence had high identity to members of CYP4,and suggested that the sequence belongs to CYP4 family.Its identity to CYP4J1,CYP4J2 and CYP4J3,the only three members of CYP4J subfamily,is 51.56%,55.49% and 56.25% respectively,which highly suggested that the sequence is one of members of CYP4J subfamily.
出处
《寄生虫与医学昆虫学报》
CAS
1998年第3期166-173,共8页
Acta Parasitologica et Medica Entomologica Sinica
基金
国家教委回国人员启动基金
广东省自然科学基金
关键词
致倦库蚊
细胞色素P450
抗药性
同源性分析
Culex quinquefasciatus\ \ Cytochrome P450\ \ Insecticide resistance\ \ Gene clone \ \ Homologous analysis
